Lentiviral-mediated gene transfer into haematopoietic stem cells
(2001) In Journal of Internal Medicine 249(4). p.339-343- Abstract
OBJECTIVES: Lentiviral vectors can transduce nondividing cells. As most haematopoietic stem cells (HSCs) are nondividing in vivo, lentiviral vectors are promising viral vectors to transfer genes into HSCs.
DESIGN AND SETTING: We have used HIV-1 based lentiviral vectors containing the green fluorescent protein (GFP) gene to transduce umbilical cord blood CD34+ and CD34+/CD38- cells prior to transplantation into NOD/SCID mice.
RESULTS: High level engraftment of human cells was obtained and transgene expression was seen in both myeloid and lymphoid lineages. Bone marrow from the primary transplant recipients mice was transplanted into secondary recipients. GFP expression was seen in both lymphoid and myeloid cells in the... (More)
OBJECTIVES: Lentiviral vectors can transduce nondividing cells. As most haematopoietic stem cells (HSCs) are nondividing in vivo, lentiviral vectors are promising viral vectors to transfer genes into HSCs.
DESIGN AND SETTING: We have used HIV-1 based lentiviral vectors containing the green fluorescent protein (GFP) gene to transduce umbilical cord blood CD34+ and CD34+/CD38- cells prior to transplantation into NOD/SCID mice.
RESULTS: High level engraftment of human cells was obtained and transgene expression was seen in both myeloid and lymphoid lineages. Bone marrow from the primary transplant recipients mice was transplanted into secondary recipients. GFP expression was seen in both lymphoid and myeloid cells in the secondary recipients 6 weeks posttransplantation. Human haematopoietic progenitor colonies were grown from both primary and secondary recipients. Over 50% of the haematopoietic colonies in these recipients were positive for the GFP transgene by PCR. Following inverse PCR, amplified fragments were sequenced and integration of the vector into human genomic DNA was demonstrated. Several vectors containing different internal promoters were tested in NOD/SCID mice that had been transplanted with transduced CD34+ and CD34+/CD38- cells. The elongation factor-1alpha (EF-1alpha) promoter gave the highest level of expression, both in the myeloid and lymphoid progeny of the engrafting cells.
CONCLUSIONS: These data collectively indicate that candidate human HSCs can be efficiently transduced with lentiviral vectors and that the transgene is highly expressed in their progeny cells.
(Less)
- author
- Woods, N B LU ; Mikkola, H ; Nilsson, E LU ; Olsson, K LU ; Trono, D and Karlsson, S
- organization
- publishing date
- 2001-04
- type
- Contribution to journal
- publication status
- published
- keywords
- Animals, Antigens, CD34, Gene Transfer Techniques, Genetic Vectors, Hematopoietic Stem Cells, Humans, Lentivirus, Mice, Mice, SCID, Promoter Regions, Genetic, Transduction, Genetic
- in
- Journal of Internal Medicine
- volume
- 249
- issue
- 4
- pages
- 5 pages
- publisher
- Wiley-Blackwell
- external identifiers
-
- pmid:11298854
- scopus:0035731632
- ISSN
- 0954-6820
- DOI
- 10.1046/j.1365-2796.2001.00806.x
- language
- English
- LU publication?
- yes
- id
- a731b6d7-23d7-4aa5-bf64-1486ed9a8daa
- date added to LUP
- 2019-11-25 13:51:03
- date last changed
- 2024-01-02 00:33:44
@article{a731b6d7-23d7-4aa5-bf64-1486ed9a8daa,
abstract = {{<p>OBJECTIVES: Lentiviral vectors can transduce nondividing cells. As most haematopoietic stem cells (HSCs) are nondividing in vivo, lentiviral vectors are promising viral vectors to transfer genes into HSCs.</p><p>DESIGN AND SETTING: We have used HIV-1 based lentiviral vectors containing the green fluorescent protein (GFP) gene to transduce umbilical cord blood CD34+ and CD34+/CD38- cells prior to transplantation into NOD/SCID mice.</p><p>RESULTS: High level engraftment of human cells was obtained and transgene expression was seen in both myeloid and lymphoid lineages. Bone marrow from the primary transplant recipients mice was transplanted into secondary recipients. GFP expression was seen in both lymphoid and myeloid cells in the secondary recipients 6 weeks posttransplantation. Human haematopoietic progenitor colonies were grown from both primary and secondary recipients. Over 50% of the haematopoietic colonies in these recipients were positive for the GFP transgene by PCR. Following inverse PCR, amplified fragments were sequenced and integration of the vector into human genomic DNA was demonstrated. Several vectors containing different internal promoters were tested in NOD/SCID mice that had been transplanted with transduced CD34+ and CD34+/CD38- cells. The elongation factor-1alpha (EF-1alpha) promoter gave the highest level of expression, both in the myeloid and lymphoid progeny of the engrafting cells.</p><p>CONCLUSIONS: These data collectively indicate that candidate human HSCs can be efficiently transduced with lentiviral vectors and that the transgene is highly expressed in their progeny cells.</p>}},
author = {{Woods, N B and Mikkola, H and Nilsson, E and Olsson, K and Trono, D and Karlsson, S}},
issn = {{0954-6820}},
keywords = {{Animals; Antigens, CD34; Gene Transfer Techniques; Genetic Vectors; Hematopoietic Stem Cells; Humans; Lentivirus; Mice; Mice, SCID; Promoter Regions, Genetic; Transduction, Genetic}},
language = {{eng}},
number = {{4}},
pages = {{339--343}},
publisher = {{Wiley-Blackwell}},
series = {{Journal of Internal Medicine}},
title = {{Lentiviral-mediated gene transfer into haematopoietic stem cells}},
url = {{http://dx.doi.org/10.1046/j.1365-2796.2001.00806.x}},
doi = {{10.1046/j.1365-2796.2001.00806.x}},
volume = {{249}},
year = {{2001}},
}